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Fig. 1. Cx43 induces the formation of functional gap junction channels in LNCaP but not PC3 cells. (A,B) Photomicrographs show the distribution of Cx43 immunoreactivity in PC3 cells that were uninfected (A) or infected with Ad-Cx43 (B). (C-F) Photomicrographs show the distribution of connexin immunoreactivity in LNCaP cells that were uninfected (C) or infected with Ad-Cx43 (D), Ad-Cx37 (E), or Ad-Cx40 (F). (G) Immunoblot detection of Cx43 in cell lysates from PC3 cells that were uninfected (lane 1) or infected with Ad-Cx43 (lane 2) and in lysates of uninfected LNCaP cells (lane 3) or LNCaP cells infected with Ad-Cx43 (lane 4) or Ad-Cx43DN (lane 5). The migration positions of the molecular mass markers are indicated on the right. (H-K) Representative fluorescence images of Lucifer Yellow transfer after microinjection of the tracer into a single cell within a cluster of uninfected LNCaP cells (H) or LNCaP cells infected with Ad-Cx43 (I), Ad-Cx43DN (J), or both Ad-Cx43 and Ad-Cx43DN (K). (L) Histogram showing the extent of intercellular transfer of Lucifer Yellow in uninfected LNCaP cells and LNCaP cells infected with Ad-Cx43, Ad-Cx43DN, or both adenoviruses. Values represent the number of recipient cells containing Lucifer Yellow (mean ± s.e.m., n=8). (M-O) Photomicrographs show the localization of Cx43DN (M) and Golgi 58K protein (N) in LNCaP cells infected with Ad-Cx43DN after double-labeling immunofluorescence using anti-Cx43 and anti-Golgi 58K antibodies. The overlap of the two signals appears yellow in the merged image (O). (P-R) Photomicrographs show the localization of Cx43 and Cx43DN in LNCaP cells co-infected with Ad-Cx43 and Ad-Cx43DN after double-labeling immunofluorescence using anti-Cx43IL (P) and anti-HA (Q) antibodies. The overlap of anti-Cx43IL and anti-HA immunoreactivities appears yellow (R). Bar, 20 µm for A-D and P-R, 8 µm for E, 12 µm for F, 66 µm for H-K and 17 µm for M-O.
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